Defining the roles of an enhancer long non-coding RNA eIncRNA-ID2 in rickettsial pathogenesis and immunity

定义增强子长链非编码 RNA eIncRNA-ID2 在立克次体发病机制和免疫中的作用

基本信息

批准号：
10323675
负责人：
Abha Sahni
金额：
$ 19.75万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-04 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10323675
关键词：
Address Affect Agreement Animal Model Anti-Infective Agents Area B-Lymphocytes Bacteria Biogenesis Biological Biological Process Biology Blood Brain C3H/HeN Mouse CD8-Positive T-Lymphocytes Cells Cellular Immunity Cerebral Edema ChIP-seq Chemosensitization Code Communicable Diseases DNA Data Set Dendritic Cells Deoxyribonuclease I Development Disease Disease Progression E protein EP300 gene Edema Elements Encyclopedia of DNA Elements Endothelial Cells Enhancers Epigenetic Process Functional disorder Gene Expression Gene Expression Regulation Generations Genes Genetic Transcription Genomics Goals Helix-Loop-Helix Motifs Human Human Genome Hypersensitivity ID2 gene Immune Immune System Diseases Immune response Immune system Immunity Immunology In Vitro Infection Inflammation Inhibitor of Differentiation Proteins Investigation Knowledge Link Lung Lysine Maintenance Mediating Messenger RNA Microbiology Modification Molecular Mus Natural Killer Cells Nucleotides Organ Outcome POLR2A gene Pathogenesis Pathogenicity Physiological Poly(A) Tail Predisposition Proteins Publishing RNA RNA Polymerase II RNA Splicing Regulation Regulator Genes Rickettsia Rickettsia Infections Rickettsia conorii Role Shapes Site Spleen System T memory cell T-Lymphocyte Testing Therapeutic Tissues Transcript Transcription Initiation Site Transcriptional Regulation Untranslated RNA Up-Regulation Vaccine Adjuvant Vascular Permeabilities alpha helix antagonist base body system cell mediated immune response cell type dark matter defined contribution follow-up histone methylation human disease human model immune activation in vivo Model insight macrophage mammalian genome microbial migration monolayer mouse model novel novel strategies pathogenic microbe polypeptide prevent programs promoter response spotted fever therapeutic development therapeutic target tick-borne transcription factor transcriptome sequencing vascular inflammation

项目摘要

PROJECT SUMMARY Precise and dynamic alterations in gene expression are critical determinants of the regulation of host immunity to microbial pathogens. Pathogenic rickettsiae in the spotted fever group cause some of the most severe infectious diseases in humans, characterized by microvascular inflammation and dysfunction attributed to disseminated infection of endothelial cells and increased vascular permeability resulting in pulmonary/cerebral edema. Long non-coding (lnc) RNAs of ≥ 200 nucleotides regulate a panoply of biological responses through an array of mechanisms and changes in their expression levels are now intricately linked to the determination of innate as well as cell-mediated immune responses. As an important subset of lncRNAs, enhancer lncRNAs implement their regulatory roles by enhancing protein coding genes (PCGs) in a cis- or trans-acting manner. We performed RNA-sequencing on the lungs as one of the predominantly affected target organs of susceptible mice infected with R. conorii to identify up-regulation of 179 lncRNAs. Via follow-up analysis to differentiate enhancer (elnc) from promoter-associated (plnc) RNAs based on the ratio of single- versus tri-methylation of histone 3 at lysine 4 (H3K4Me1:H3K4Me3) and other active enhancer signatures based on POLR2A, p300, DNase I hypersensitivity sites, CTCF, and Hi-3C ChIP-Seq datasets, we further determined significantly higher expression of an active elncRNA013718 and its target PCG Inhibitor of DNA binding 2 (ID2) in the mouse lungs, spleen, and CD8+ T-cells during Rickettsia conorii infection. Our preliminary findings further suggest that elncRNA013718 positively regulates the expression of ID2, a protein antagonist of E protein transcription factors and a regulator of T cells in the immune system. Accordingly, we refer to elncRNA013718 as elncRNA- ID2 and hypothesize novel contributory roles for elncRNA-ID2:Id2 interplay in the regulation of protective host immunity during rickettsial infections. We propose to test this hypothesis via two independent yet thematically interlinked specific aims. Aim 1 will distinguish cell type-specific expression and functional roles of elncRNA- ID2 and ID2 in the host lungs and spleen in experimental murine models of R. conorii and R. australis infection. In Aim 2, we will determine the modulatory effects of both global and cell-specific interference with elncRNA- ID2 on host immune responses and disease progression/outcome. Given the complexity of cellular immune responses, we will employ two independent and established in vivo models of infection closely mimicking the pathophysiology of human rickettsioses and cutting-edge approaches of cellular microbiology and immunology to determine the regulatory potential of elncRNA-ID2 as a novel elncRNA in the molecular circuitry underlying regulation of host immunity. The acquired insights will enhance our knowledge of context-specific physiological roles of elncRNA-ID2 in the determination of host responses to pathogenic rickettsiae and reveal potentially unique entry points for novel strategies to enhance host immunity against intracellular microbial infections.

项目概要基因表达的精确和动态改变是宿主免疫调节的关键决定因素斑点热病群中的致病性立克次体会引起一些最严重的症状。人类传染病，其特征是微血管炎症和功能障碍内皮细胞的播散性感染和血管通透性增加导致肺/脑损伤 ≥ 200 个核苷酸的长非编码 (lnc) RNA 通过调节一系列生物反应。一系列机制及其表达水平的变化现在与该决定错综复杂地联系在一起作为 lncRNA 的重要子集，增强子 lncRNA 参与了先天性和细胞介导的免疫反应。通过以顺式或反式作用方式增强蛋白质编码基因（PCG）来发挥其调节作用。我们对肺部进行了 RNA 测序，这是主要受影响的易感性靶器官之一感染 R. conorii 的小鼠通过后续分析来鉴定 179 个 lncRNA 的上调。启动子相关 (plnc) RNA 的增强子 (elnc) 基于单甲基化与三甲基化的比率赖氨酸 4 处的组蛋白 3 (H3K4Me1:H3K4Me3) 和其他基于 POLR2A、p300、 DNase I 超敏位点、CTCF 和 Hi-3C ChIP-Seq 数据集，我们进一步确定显着更高活性 elncRNA013718 及其靶标 PCG DNA 结合抑制剂 2 (ID2) 在小鼠中的表达我们的初步研究结果进一步表明，康氏立克次体感染期间肺、脾和 CD8+ T 细胞。 elncRNA013718 正向调节 ID2（E 蛋白转录的蛋白拮抗剂）的表达因此，我们将 elncRNA013718 称为 elncRNA-。 ID2 和 elncRNA-ID2 的新贡献作用：Id2 在保护性宿主调节中相互作用我们建议通过两个独立但主题性的方法来检验这一假设。目标 1 将区分 elncRNA 的细胞类型特异性表达和功能作用。 R. conorii 和 R. australis 感染的实验鼠模型中宿主肺和脾中的 ID2 和 ID2。在目标 2 中，我们将确定 elncRNA 的全局和细胞特异性干扰的调节作用鉴于细胞免疫的复杂性，ID2 对宿主免疫反应和疾病进展/结果的影响。响应，我们将采用两个独立且已建立的体内感染模型，密切模仿人类立克次体病的病理生理学以及细胞微生物学和免疫学的前沿方法确定 elncRNA-ID2 作为新型 elncRNA 在分子电路中的调节潜力宿主免疫的调节所获得的见解将增强我们对特定环境生理学的了解。 elncRNA-ID2 在确定宿主对致病性立克次体反应中的作用并可能揭示增强宿主对细胞内微生物感染的免疫力的新策略的独特切入点。